Method Development, Validation, and Forced Degradation Studies of Pacritinib Using HPLC and LC-MS/MS for Impurity Characterisation
نویسندگان
1 Department of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, India
2 Department of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, India
3 Analytical Research and Development, Amneal Pharmaceuticals, Piscataway, New Jersey, USA
4 Department of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, India
5 Department of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, India
doi
10.48309/ajca.2026.575116.2054چکیده
An uncomplicated, effective, and stability-indicating reverse-phase HPLC method was developed and validated for the quantitative determination of Pacritinib and its related impurities, along with the characterization of degradation products using LC–MS/MS. Chromatographic separation was achieved on a Zorbax C18 column (150 × 4.6 mm, 3.5 µm) using a gradient mobile phase consisting of acetonitrile and ammonium formate buffer (pH 2.5). Detection was carried out at 257 nm with a total run time of 24 minutes. The optimized chromatographic conditions enabled efficient separation of Pacritinib and five specified impurities with acceptable system suitability parameters. The method was validated according to ICH Q2(R1) guidelines. Pacritinib exhibited excellent linearity over the concentration range of 25–150 µg/mL (R² = 0.9999), while impurities showed linear responses within their respective ranges (R² ≥ 0.9997). Precision studies demonstrated %RSD values below 1.0% for both system and method precision. Accuracy results showed mean recoveries ranging from 99.1% to 100.4%, indicating good reliability of the method. Robustness evaluation confirmed that the method remained unaffected by small variations in flow rate and organic phase composition. Forced degradation studies under acidic, alkaline, oxidative, reductive, thermal, photolytic, and hydrolytic conditions revealed significant degradation in acid, base, and oxidative environments. Four major degradation products (DP1–DP4) were detected and characterized by LC–MS/MS based on accurate mass measurements and fragmentation patterns. The developed method is suitable for stability studies, impurity profiling, and quality control of Pacritinib in pharmaceutical formulations.
